Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes.
Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes.
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DOI：
10.1038/s41467-021-23239-0
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发表时间：
2021-05-20
影响因子：
16.6
通讯作者：
Gantz VM
中科院分区：
文献类型：
--
作者：
Feng X;López Del Amo V;Mameli E;Lee M;Bishop AL;Perrimon N;Gantz VM
库蚊是多种人类和动物疾病的全球媒介，包括西尼罗河病毒、淋巴丝虫病和禽疟疾，对公共卫生、牲畜、伴侣动物和濒危鸟类构成持续威胁。虽然杀虫剂耐药性的上升威胁到了库蚊的控制，但CRISPR基因组编辑工具的进步促进了替代遗传策略的发展，如基因驱动系统来对抗疾病载体。然而，尽管基因驱动技术在其他蚊子身上取得了迅速进展，但在库蚊身上却缺乏进展。在这里，我们开发了一个库蚊特异性Cas9/gRNA表达工具包，并使用定点同源转基因来产生和验证致倦库蚊Cas9表达系。我们发现，gRNA支架变体提高了致倦库蚊和黑腹果蝇的转基因效率，并提高了果蝇的基因驱动性能。这些发现支持了未来控制库蚊的技术开发，并为在其他物种中改进这些工具提供了有价值的见解。库蚊是昆虫传播疾病的全球性载体，尽管遗传工具的进展落后于其他蚊子物种。在这里，作者提出了一个基于Cas9的工具包和方法，可以支持这些蚊子未来的基因驱动开发。
Culex mosquitoes are a global vector for multiple human and animal diseases, including West Nile virus, lymphatic filariasis, and avian malaria, posing a constant threat to public health, livestock, companion animals, and endangered birds. While rising insecticide resistance has threatened the control of Culex mosquitoes, advances in CRISPR genome-editing tools have fostered the development of alternative genetic strategies such as gene drive systems to fight disease vectors. However, though gene-drive technology has quickly progressed in other mosquitoes, advances have been lacking in Culex. Here, we develop a Culex-specific Cas9/gRNA expression toolkit and use site-directed homology-based transgenesis to generate and validate a Culex quinquefasciatus Cas9-expressing line. We show that gRNA scaffold variants improve transgenesis efficiency in both Culex quinquefasciatus and Drosophila melanogaster and boost gene-drive performance in the fruit fly. These findings support future technology development to control Culex mosquitoes and provide valuable insight for improving these tools in other species. Culex mosquitoes are a global vector for insect-borne diseases, though progress with genetic tools lags behind other mosquito species. Here the authors present a Cas9-based toolkit and methods that could support future gene drive development in these mosquitoes.
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影响因子：
12.3
作者：
Dang Y;Jia G;Choi J;Ma H;Anaya E;Ye C;Shankar P;Wu H
通讯作者：
Wu H
DOI：
10.1126/science.aaa5945
发表时间：
2015-04-24
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Gantz VM;Bier E
通讯作者：
Bier E
影响因子：
2.1
作者：
HSU, SH;MAO, WH;CROSS, JH
通讯作者：
CROSS, JH
影响因子：
11.6
作者：
Atkinson, Carter T.;Utzurrum, Ruth B.;Giambelluca, Thomas W.
通讯作者：
Giambelluca, Thomas W.
影响因子：
1.9
作者：
Allen, ML;Christensen, BM
通讯作者：
Christensen, BM